DDAH1 effects on endogenous NOS inhibitors and vascular endothelial function

DDAH1 对内源性 NOS 抑制剂和血管内皮功能的影响

• 批准号: 8111967
• 负责人: YINGJIE CHEN
• 金额: $ 15.1万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8111967
• 关键词: Area; Arginine; Atherosclerosis; Biological Availability; Blood Vessels; Cardiovascular Diseases; Cell physiology; Clinical; Congestive Heart Failure; Coronary Arteriosclerosis; Data; Diabetes Mellitus; Endothelial Cells; Enzymes; Excretory function; Functional disorder; Gene Deletion; Gene Silencing; Genes; Goals; Hypertension; In Vitro; Kidney; Laboratories; Modeling; Molecular; Mouse Strains; Mus; Nitric Oxide; Nitric Oxide Synthase; Pathway interactions; Plasma; Play; Production; Protein Isoforms; Regulation; Relative (related person); Reporting; Research Personnel; Risk Factors; Role; Stroke; Techniques; Testing; Tissues; Vascular Endothelial Cell; angiogenesis; base; cell growth; cell injury; dimethylargininase; experience; in vivo; inhibitor/antagonist; injury and repair; innovation; insight; novel; omega-N-Methylarginine; overexpression; public health relevance

DESCRIPTION (provided by applicant): Accumulation of the endogenous NO synthase (NOS) inhibitors asymmetric dimethyl arginine (ADMA) and Ng-monomethyl-L-arginine (L-NMMA) is a major risk factor for cardiovascular diseases including hypertension, coronary artery disease, stroke, diabetes and atherosclerosis. These endogenous NOS inhibitors compete with L-arginine to inhibit NO production by NOS. ADMA and L-NMMA are eliminated principally by dimethylarginine dimethylaminohydrolase (DDAH) with a small contribution from renal excretion. It is reported that ADMA and L-NMMA are degraded by both DDAH1 and DDAH2, but the relative contributions of DDAH1 and DDAH2 have been controversial. Furthermore, whether DDAH1 can exert effects on vascular function beyond regulation of NO production is unknown. Using novel tissue specific DDAH1 deficient mice and global DDAH1 gene deficient mice generated in our laboratory, our preliminary data demonstrate that DDAH1 plays an essential role in degrading the endogenous NOS inhibitors, and that DDAH1 distributed in endothelial cells plays an important role in regulating vascular endothelial NO production and endothelial cell growth and injury repair. Most interestingly, our preliminary data further demonstrated that DDAH1 also regulates endothelial cell function through a novel molecular pathway independent of degradation the of endogenous NOS inhibitors. Based on these new findings, we propose to test two hypotheses. First, we hypothesize that DDAH1 is the principal enzyme responsible for degrading ADMA and L-NMMA. Second, we hypothesize that DDAH1 exerts additional actions beyond its role in degrading ADMA and L-NMMA that regulate vascular endothelial cell growth and injury repair. To test these hypotheses, we have generated a novel tissue specific endothelial-DDAH1 KO (endo-DDAH1 KO) mouse strain and a global DDAH1 KO mouse strain. The proposed studies will utilize in vitro and in vivo approaches to provide new insight into the function of the endogenous NOS inhibitors, as well as novel molecular mechanisms by which DDAH1 acts to maintain vascular endothelial cell growth and injury repair. PUBLIC HEALTH RELEVANCE: Accumulation of the endogenous nitric oxide synthase (NOS) inhibitors asymmetric dimethyl arginine (ADMA) and Ng-monomethyl-L-arginine (L-NMMA) is a major risk factor for cardiovascular diseases including hypertension, coronary artery disease, stroke, diabetes and atherosclerosis. DDAH1 degrades ADMA and L-NMMA to increase nitric oxide production. The proposed studies will integrate in vitro and in vivo approaches to provide new insight into the function of the endogenous NOS inhibitors, as well as novel molecular mechanisms by which DDAH1 acts to maintain vascular endothelial cell growth and injury repair.

描述（由申请人提供）：内源性一氧化氮合酶（NOS）抑制剂不对称二甲基精氨酸（ADMA）和Ng-单甲基-L-精氨酸（L-NMMA）的积累是心血管疾病（包括高血压、冠状动脉疾病）的主要危险因素、中风、糖尿病和动脉粥样硬化。这些内源性 NOS 抑制剂与 L-精氨酸竞争，抑制 NOS 产生 NO。 ADMA 和 L-NMMA 主要通过二甲基精氨酸二甲氨基水解酶 (DDAH) 消除，肾排泄也有少量贡献。据报道，ADMA和L-NMMA均被DDAH1和DDAH2降解，但DDAH1和DDAH2的相对贡献一直存在争议。此外，DDAH1 是否可以对 NO 产生的调节之外的血管功能产生影响尚不清楚。利用本实验室制备的新型组织特异性DDAH1缺陷小鼠和全局DDAH1基因缺陷小鼠，我们的初步数据表明DDAH1在降解内源性NOS抑制剂中发挥重要作用，并且分布在内皮细胞中的DDAH1在调节血管内皮细胞中发挥重要作用。 NO 产生以及内皮细胞生长和损伤修复。最有趣的是，我们的初步数据进一步表明，DDAH1 还通过一种独立于内源性 NOS 抑制剂降解的新型分子途径调节内皮细胞功能。基于这些新发现，我们建议检验两个假设。首先，我们假设 DDAH1 是负责降解 ADMA 和 L-NMMA 的主要酶。其次，我们假设 DDAH1 除了降解调节血管内皮细胞生长和损伤修复的 ADMA 和 L-NMMA 作用外，还发挥其他作用。为了检验这些假设，我们生成了一种新型组织特异性内皮-DDAH1 KO (endo-DDAH1 KO) 小鼠品系和一种全局 DDAH1 KO 小鼠品系。拟议的研究将利用体外和体内方法来提供对内源性 NOS 抑制剂功能的新见解，以及 DDAH1 维持血管内皮细胞生长和损伤修复的新分子机制。 公共健康相关性：内源性一氧化氮合酶 (NOS) 抑制剂不对称二甲基精氨酸 (ADMA) 和 Ng-单甲基-L-精氨酸 (L-NMMA) 的积累是心血管疾病的主要危险因素，包括高血压、冠状动脉疾病、中风、糖尿病和动脉粥样硬化。 DDAH1 降解 ADMA 和 L-NMMA 以增加一氧化氮的产生。拟议的研究将整合体外和体内方法，为内源性 NOS 抑制剂的功能提供新的见解，以及 DDAH1 维持血管内皮细胞生长和损伤修复的新分子机制。